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Identification and characterization of a HEPN-MNT family type II toxin–antitoxin in Shewanella oneidensis

机译:一株希瓦氏假单胞菌HEPN-MNT家族II型毒素-抗毒素的鉴定和表征

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摘要

Toxin–antitoxin (TA) systems are prevalent in bacteria and archaea. However, related studies in the ecologically and bioelectrochemically important strain Shewanella oneidensis are limited. Here, we show that SO_3166, a member of the higher eukaryotes and prokaryotes nucleotide-binding (HEPN) superfamily, strongly inhibited cell growth in S. oneidensis and Escherichia coli. SO_3165, a putative minimal nucleotidyltransferase (MNT), neutralized the toxicity of SO_3166. Gene SO_3165 lies upstream of SO_3166, and they are co-transcribed. Moreover, the SO_3165 and SO_3166 proteins interact with each other directly in vivo, and antitoxin SO_3165 bound to the promoter of the TA operon and repressed its activity. Finally, the conserved Rx4-6H domain in HEPN family was identified in SO_3166. Mutating either the R or H abolished SO_3166 toxicity, confirming that Rx4-6H domain is critical for SO_3166 activity. Taken together, these results demonstrate that SO_3166 and SO_3165 in S. oneidensis form a typical type II TA pair. This TA pair plays a critical role in regulating bacterial functions because its disruption led to impaired cell motility in S. oneidensis. Thus, we demonstrated for the first time that HEPN-MNT can function as a TA system, thereby providing important insights into the understanding of the function and regulation of HEPNs and MNTs in prokaryotes.
机译:毒素-抗毒素(TA)系统在细菌和古细菌中很普遍。但是,对生态和生物电化学重要菌株希瓦氏菌(Shewanella oneidensis)的相关研究是有限的。在这里,我们显示SO_3166,高等真核生物和原核生物核苷酸结合(HEPN)超家族的成员,强烈抑制oneidensis和大肠杆菌中的细胞生长。推定的最小核苷酸转移酶(MNT)SO_3165中和了SO_3166的毒性。基因SO_3165位于SO_3166的上游,并且它们被共转录。此外,SO_3165和SO_3166蛋白在体内直接相互作用,抗毒素SO_3165与TA操纵子的启动子结合并抑制其活性。最后,在SO_3166中鉴定了HEPN家族中的保守Rx4-6H结构域。突变R或H消除了SO_3166毒性,证实Rx4-6H结构域对于SO_3166活性至关重要。总之,这些结果表明,沙门氏菌中的SO_3166和SO_3165形成典型的II型TA对。该TA对在调节细菌功能中起关键作用,因为它的破坏导致S. oneidensis的细胞运动受损。因此,我们首次证明了HEPN-MNT可以作为TA系统发挥作用,从而为了解原核生物中HEPNs和MNT的功能和调控提供了重要的见识。

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